Ultrafast MAS Solid-State NMR Permits Extensive 13C and 1H Detection in Paramagnetic Metalloproteins
Ultrafast MAS Solid-State NMR Permits Extensive 13C and 1H Detection in Paramagnetic Metalloproteins
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DOI：
10.1021/ja100398q
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发表时间：
2010-04-28
影响因子：
15
通讯作者：
Pintacuda, Guido
中科院分区：
文献类型：
--
作者：
Bertini, Ivano;Emsley, Lyndon;Pintacuda, Guido
我们在这里展示了结合基于超快（60 kHz） MAS的定制方法，在co - ii取代的基质金属蛋白酶12 （comp -12）的催化区域上，我们可以观察和分配，在固态的高顺磁性蛋白质中，协调金属中心的残基的C-13甚至H-1共振。此外，通过利用顺磁中心引起的增强弛豫，以及快速MAS实现的低功率辐照，这可以在非常短的时间和非常高的场（21.2 T）下实现，只有不到1 mg的样品。此外，利用已知的化合物晶体结构，我们能够区分和测量假接触（PCS）对配位配体位移的贡献，并揭示结构信息。
We show here that by combining tailored approaches based on ultrafast (60 kHz) MAS on the Co-II-replaced catalytic domain of matrix metalloproteinase 12 (CoMMP-12) we can observe and assign, in a highly paramagnetic protein in the solid state, C-13 and even H-1 resonances from the residues coordinating the metal center. In addition, by exploiting the enhanced relaxation caused by the paramagnetic center, and the low power irradiation enabled by the fast MAS, this can be achieved in remarkably short times and at very high field (21.2 T), with only less than 1 mg of sample. Furthermore, using the known crystal structure of the compound, we are able to distinguish and measure pseudocontact (PCS) contributions to the shifts up to the coordinating ligands and to unveil structural information.